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Kimiko HONSHO Michiko HIROSE Masanori HATORI Lubna YASMIN Haruna IZU Shogo MATOBA Sumie TOGAYACHI Hiroyuki MIYOSHI Tadashi SANKAI Atsuo OGURA Arata HONDA 《The Journal of reproduction and development》2015,61(1):13-19
Quality evaluation of pluripotent stem cells using appropriate animal models needs to be improved for human regenerative medicine. Previously, we demonstrated that although the in vitro neural differentiating capacity of rabbit induced pluripotent stem cells (iPSCs) can be mitigated by improving their baseline level of pluripotency, i.e., by converting them into the so-called “naïve-like” state, the effect after such conversion of rabbit embryonic stem cells (ESCs) remains to be elucidated. Here we found that naïve-like conversion enhanced the differences in innate in vitro differentiation capacity between ESCs and iPSCs. Naïve-like rabbit ESCs exhibited several features indicating pluripotency, including the capacity for teratoma formation. They differentiated into mature oligodendrocytes much more effectively (3.3–7.2 times) than naïve-like iPSCs. This suggests an inherent variation in differentiation potential in
vitro among PSC lines. When naïve-like ESCs were injected into preimplantation rabbit embryos, although they contributed efficiently to forming the inner cell mass of blastocysts, no chimeric pups were obtained. Thus, in vitro neural differentiation following naïve-like conversion is a promising option for determining the quality of PSCs without the need to demonstrate chimeric contribution. These results provide an opportunity to evaluate which pluripotent stem cells or treatments are best suited for therapeutic use. 相似文献
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Mami OIKAWA Shogo MATOBA Kimiko INOUE Satoshi KAMIMURA Michiko HIROSE Narumi OGONUKI Hirosuke SHIURA Michihiko SUGIMOTO Kuniya ABE Fumitoshi ISHINO Atsuo OGURA 《The Journal of reproduction and development》2013,59(3):231-237
In mice, one of the major epigenetic errors associated with somatic cell nuclear
transfer (SCNT) is ectopic expression of Xist during the preimplantation
period in both sexes. We found that this aberrant Xist expression could
be impeded by deletion of Xist from the putative active X chromosome in
donor cells. In male clones, it was also found that prior injection of
Xist-specific siRNA could significantly improve the postimplantation
development of cloned embryos as a result of a significant repression of
Xist at the morula stage. In this study, we examined whether the same
knockdown strategy could work as well in female SCNT-derived embryos. Embryos were
reconstructed with cumulus cell nuclei and injected with Xist-specific
siRNA at 6–7 h after oocyte activation. RNA FISH analysis revealed that siRNA treatment
successfully repressed Xist RNA at the morula stage, as shown by the
significant decrease in the number of cloud-type Xist signals in the
blastomere nuclei. However, blastomeres with different sizes (from “pinpoint” to “cloud”)
and numbers of Xist RNA signals remained within single embryos. After
implantation, the dysregulated Xist expression was normalized
autonomously, as in male clones, to a state of monoallelic expression in both embryonic
and extraembryonic tissues. However, at term there was no significant improvement in the
survival of the siRNA-injected cloned embryos. Thus, siRNA injection was largely effective
in repressing the Xist overexpression in female cloned embryos but failed
to rescue them, probably because of an inability to mimic consistent monoallelic
Xist expression in these embryos. This could only be achieved in female
embryos by applying a gene knockout strategy rather than an siRNA approach. 相似文献
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Kiyoshi AKIYAMA Jun KOBAYASHI Yoshimasa SATO Ryuichi SATA Megumi OHASHI Emi SASAKI Yorimasa ODA Yoshio OGAWA Shuji UEDA Hisashi NABENISHI Satoko MATOBA 《Animal Science Journal》2010,81(4):461-466
The objective of this study was to develop an in‐straw dilution method suitable for direct transfer of vitrified bovine sexed embryos. Embryo sexing was performed by molecular diagnosis. Several sexed and vitrified‐warmed embryos were transferred after evaluation of morphologically embryonic survival at warming and in‐straw dilution (Evaluation group). The other embryos were immediately directly transferred to recipients without first being expelled from the straws after in‐straw dilution (Non‐evaluation group). The pregnancy rates of vitrified sexed embryos were 38.7% and 34.8% in the Evaluation group and Non‐evaluation group, respectively, which were not significantly different. The viability of lower quality embryos before vitrification tended to be lower (P = 0.087) than that of the higher quality embryos regardless of evaluating embryos after warming and in‐straw dilution. The abortion rates were similar, and there was no difference between the two groups (13.9% and 12.5%, respectively). These results demonstrate that vitrified bovine sexed embryos can be vitrified and diluted by the in‐straw method and that the vitrified and warmed sexed embryos can develop to term. 相似文献
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